# Use quartz cuvette specially made for UV-Vis and fill cuvette ¾ full with HPLC grade H<sub>2</sub>O

# Use quartz cuvette specially made for UV-Vis and fill cuvette ¾ full with HPLC grade H<sub>2</sub>O

Line 30:

Line 44:

# Run baseline of blank sample (HPLC grade H<sub>2</sub>O)

# Run baseline of blank sample (HPLC grade H<sub>2</sub>O)

# Once finished running, autoscale the y-axis on the screen until max absorption = 1.0

# Once finished running, autoscale the y-axis on the screen until max absorption = 1.0

-

# Save sample name under new folder MKM

+

# Save sample name under new folder '''MKM'''

'''Repeat procedure for all UV-Vis Samples run; however, do not have to run a baseline every time after each sample. Baseline only has to be run once throughout the total analysis. Also please be sure to clean each cuvette and wipe thoroughly each time before running new samples for analysis. In addition, place actual samples that are not references in the first pocket of six sample holder in UV-Vis.'''

'''Repeat procedure for all UV-Vis Samples run; however, do not have to run a baseline every time after each sample. Baseline only has to be run once throughout the total analysis. Also please be sure to clean each cuvette and wipe thoroughly each time before running new samples for analysis. In addition, place actual samples that are not references in the first pocket of six sample holder in UV-Vis.'''

+

+

==Porphyrin Dilution Calculations==

+

+

'''STOCK SOLUTION PORPHYRIN STARTED WITH CONCENTRATION OF 94.45μM, DILUTE SOLUTION TO GIVEN CONCENTRATIONS LISTED'''

* The concentrations of 31.48μM and 62.97μM proved to be too high in concentration; absorbency values were greater than 1.0 for each solution.

==Porphyrin Film Preparations==

==Porphyrin Film Preparations==

-

'''Follow procedural protocol for preparing glutaldehyde films 9/25/12; however, add ~ 1mL of porphyrin in addition to glutaldehyde for each film before pouring substance into a Teflon dish.'''

+

'''Follow procedural protocol for preparing glutaldehyde films 9/19/12; however, add ~ 1mL of porphyrin in addition to glutaldehyde to each film before pouring substance into a Teflon dish. Also for two PVOH films, be sure to add two drops of HCl to observe if the addition of the acid aids in securing all of the porphyrin within the films.'''

Use quartz cuvette specially made for UV-Vis and fill cuvette ¾ full with HPLC grade H2O

Wipe quartz cuvette gently and thoroughly with a Kim Wipe

Place cuvette in reference pocket of six sample UV-Vis instrument

Approve selection and file name then click baseline button

Run baseline of blank sample (HPLC grade H2O)

Once finished running, autoscale the y-axis on the screen until max absorption = 1.0

Save sample name under new folder MKM

Repeat procedure for all UV-Vis Samples run; however, do not have to run a baseline every time after each sample. Baseline only has to be run once throughout the total analysis. Also please be sure to clean each cuvette and wipe thoroughly each time before running new samples for analysis. In addition, place actual samples that are not references in the first pocket of six sample holder in UV-Vis.

Porphyrin Dilution Calculations

STOCK SOLUTION PORPHYRIN STARTED WITH CONCENTRATION OF 94.45μM, DILUTE SOLUTION TO GIVEN CONCENTRATIONS LISTED

Solution of 9.445 μM porphyrin

(94.45 μM)(V1)=(9.445 μM)(10mL)
V1 = 1mL of 94.45 μM porphyrin

Solution of 944.5 nM porphyrin

(9.445 μM)(V1)=(944.5 nM)(10mL)
V1 = 1mL of 9.445 μM porphyrin

Solution of 94.45 nM porphyrin

(944.5 nM)(V1)=(94.45 nM)(10mL)
V1 = 1mL of 944.5 nM porphyrin

Solution of 9.445 nM porphyrin

(944.5 nM)(V1)=(9.445 nM)(10mL)
V1 = 0.1mL of 944.5 nM porphyrin

Solution of 1.89 μM porphyrin

(94.45 μM)(V1)=(1.89 μM)(10mL)
V1 = 0.2mL of 94.45 μM porphyrin

Solution of 3.78 uM porphyrin

(94.45 μM)(V1)=(3.78 μM)(10mL)
V1 = 0.4mL of 94.45 μM porphyrin

UV-Vis of Porphyrin Solutions

Using a Shimadzu UV-2550 spectrophotometer, spectra of four concentrations of porphyrin solutions were taken:

* Wavelength range = 200 - 800nm
* Slit width = 1.0nm

UV-Vis Analysis Notes

The baseline was conducted with nothing in the cell.

For solutions at concentrations 944.5nM and 94.45nM, two peaks were visible at 410nm and 430nm.

The 9.445nM was not concentrated enough to produce any peaks.

The 944.5nM solution displayed several more peaks past 500nm, producing more useful data.

The concentrations of 31.48μM and 62.97μM proved to be too high in concentration; absorbency values were greater than 1.0 for each solution.

Porphyrin Film Preparations

Follow procedural protocol for preparing glutaldehyde films 9/19/12; however, add ~ 1mL of porphyrin in addition to glutaldehyde to each film before pouring substance into a Teflon dish. Also for two PVOH films, be sure to add two drops of HCl to observe if the addition of the acid aids in securing all of the porphyrin within the films.